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Updated: Jan 1, 2026
Characterization of Immune Cell-derived Extracellular Vesicles and Studying Functional Impact on Cell Environment
Published on: June 2, 2020
Effects of Pseudomonas aeruginosa on Microglial-Derived Extracellular Vesicle Biogenesis and Composition
Leandra B Jones1, Sanjay Kumar2, Courtnee' R Bell1
1Microbiology Program, Department of Biological Sciences, College of Science, Technology, Engineering and Mathematics, Alabama State University, Montgomery, AL 36104, USA.
Abstract:
The packaging of molecular constituents inside extracellular vesicles (EVs) allows them to participate in intercellular communication and the transfer of biological molecules, however the role of EVs during bacterial infection is poorly understood. The goal of this study was to examine the effects of Pseudomonas aeruginosa (P. aeruginosa) infection on the biogenesis and composition of EVs derived from the mouse microglia cell line, BV-2. BV-2 cells were cultured in exosome-free media and infected with 0, 1.3 × 104, or 2.6 × 104 colony forming units per milliliter P. aeruginosa for 72 h. The results indicated that compared with the control group, BV-2 cell viability significantly decreased after P. aeruginosa infection and BV-2-derived EVs concentration decreased significantly in the P. aeruginosa-infected group. P. aeruginosa infection significantly decreased chemokine ligand 4 messenger RNA in BV-2-derived infected EVs, compared with the control group (p ≤ 0.05). This study also revealed that heat shock protein 70 (p ≤ 0.05) and heat shock protein 90β (p ≤ 0.001) levels of expression within EVs increased after P. aeruginosa infection. EV treatment with EVs derived from P. aeruginosa infection reduced cell viability of BV-2 cells. P. aeruginosa infection alters the expression of specific proteins and mRNA in EVs. Our study suggests that P. aeruginosa infection modulates EV biogenesis and composition, which may influence bacterial pathogenesis and infection.
Insights
Pseudomonas aeruginosa infection impacts extracellular vesicle (EV) production and content in mouse microglia. These altered EVs, containing increased heat shock proteins, reduce cell viability, suggesting a role in bacterial pathogenesis.
Area of Science:
- Microbiology and Immunology
- Cell Biology
- Extracellular Vesicle Research
Background:
- Extracellular vesicles (EVs) mediate intercellular communication by transferring molecular cargo.
- The role of EVs in bacterial infections, particularly Pseudomonas aeruginosa, is not well understood.
- Microglia are key immune cells in the central nervous system, responding to pathogens.
Purpose of the Study:
- To investigate the effects of Pseudomonas aeruginosa infection on the biogenesis and composition of EVs from mouse BV-2 microglia cells.
- To determine how P. aeruginosa infection alters EV production and molecular cargo.
- To assess the impact of P. aeruginosa-infected EVs on microglia cell viability.
Main Methods:
- BV-2 microglia cells were cultured and infected with varying concentrations of P. aeruginosa for 72 hours.
- EVs were isolated from cell culture supernatant.
- EV concentration, cell viability, mRNA (chemokine ligand 4), and protein levels (heat shock proteins 70 and 90β) in EVs were analyzed.
Main Results:
- P. aeruginosa infection significantly decreased BV-2 cell viability and the concentration of derived EVs.
- Infected EVs showed significantly decreased chemokine ligand 4 mRNA.
- Levels of heat shock protein 70 and heat shock protein 90β were significantly increased in EVs from infected cells.
- Treatment with EVs from P. aeruginosa-infected cells reduced BV-2 cell viability.
Conclusions:
- Pseudomonas aeruginosa infection alters EV biogenesis and composition in microglia.
- Changes in EV cargo, including increased heat shock proteins and altered mRNA, may contribute to disease progression.
- Modulated EVs play a role in the host-pathogen interaction during P. aeruginosa infection.
